Method for animal tissue material disinfection and sterilization and corresponding animal tissue soak solution

Abstract

A method for disinfecting and sterilizing an animal tissue material and a corresponding soak solution for an animal tissue. The method includes placing the animal tissue material into an alkaline soak solution containing a metal peroxide and a detergent, and shaking; removing organic components released from microorganisms and animal tissue cells by soaking and washing in a neutral cleaning solution; washing the tissue matrix with a weak acidic cleaning solution; cryopreserving or freeze-drying the tissue matrix in a neutral solution. The soak solution for the animal tissue contains 0.01˜0.2% (w/v) of the metal peroxide and 0.05˜1.0% (w/v) of the detergent. The animal tissue pretreated by the method is advantageous to the preservation and further decellularization treatment of the tissue for manufacture of an intact scaffold material of a tissue matrix.

What we claim is: 
     
       1. A method for disinfecting and sterilizing an animal tissue, comprising the steps of:
 (a) placing the animal tissue in an alkaline soak solution containing a metal peroxide and a detergent, and shaking; 
 (b) washing the animal tissue obtained in step (a) with a neutral cleaning solution to remove organic components released from microorganisms and animal tissue cells during soaking, to obtain an animal tissue matrix; 
 (c) washing the animal tissue matrix obtained in step (b) with a weak acidic cleaning solution; 
 (d) cryopreserving or freeze-drying the animal tissue matrix obtained in step (c) in a neutral solution; 
 wherein the animal tissue is dermis tissue, and wherein the alkaline soak solution in step (a) contains 0.01˜0.2% w/v of the metal peroxide and 0.05˜1.0% w/v of the detergent, and the pH of the soak solution is between 9.5˜11.5. 
 
     
     
       2. The method according to  claim 1 , wherein the metal peroxide is selected from one or more of the group consisting of calcium peroxide, magnesium peroxide, sodium peroxide or barium peroxide, and the detergent is selected from one or more of the group consisting of polyethylene glycol octylphenol ether, sodium deoxycholate or sodium dodecyl sulfonate. 
     
     
       3. The method according to  claim 1 , wherein the step (a) is to soak each kilogram of the animal tissue in 2˜8 liter of the alkaline soak solution at the temperature of 5˜42° C. for 6˜48 hours, and shake during the soaking. 
     
     
       4. The method according to  claim 1 , wherein the neutral cleaning solution in step (b) is pure water, a saline or a biological buffer solution which is obtained by sterile filtration. 
     
     
       5. The method according to  claim 1 , wherein the animal tissue in step (b) is washed with the neutral cleaning solution for 2˜5 times, each time for 1˜3 hours. 
     
     
       6. The method according to  claim 1 , wherein the weak acidic cleaning solution in step (c) is a saline or a buffer solution of 2˜20 mM of hydroxyethylpiperazine ethane sulfonic acid, the saline or the buffer solution is obtained by adjusting pH to 5.0-6.0 with acetic acid, sodium acetate or hydrochloric acid and by sterile filtration. 
     
     
       7. The method according to  claim 1 , wherein the animal tissue matrix in step (c) is washed with the weak acidic cleaning solution for 2-5 times, each time for 1˜3 hours. 
     
     
       8. The method according to  claim 1 , wherein the neutral solution in step (d) is a buffer solution of 2˜20 mM of hydroxyethylpiperazine ethane sulfonic acid with a pH between 7.0˜8.0. 
     
     
       9. The method according to  claim 1 , wherein the alkaline soak solution further comprises a protease inhibitor which is selected from phenylmethylsulfonyl fluoride and/or N-ethylmaleimide. 
     
     
       10. The method according to  claim 2 , wherein the step (a) is to soak each kilogram of the animal tissue in 2˜8 liter of the alkaline soak solution at the temperature of 5˜42° C. for 6˜48 hours, and shake during the soaking.